1. Volume 135, Issue 6, Pages 2107-2118 (December 2008)
Identification of CD4 T-Cell Epitopes in Soluble Liver Antigen/Liver Pancreas Autoantigen in Autoimmune Hepatitis 
Heiko Mix, Christina Weiler–Normann, Robert Thimme, Golo Ahlenstiel, Eui–Cheol Shin, Johannes Herkel, Chella S. David, Ansgar W. Lohse, Barbara Rehermann 
Gastroenterology 
Volume 135, Issue 6, Pages (December 2008)
DOI: /j.gastro
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2. Figure 1 Experimental strategy. β-gal, β-galactosidase.
Gastroenterology  , DOI: ( /j.gastro )
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3. Figure 2
Humoral immune response of SLA/LP-immunized HLA-DRB1*0301–transgenic mice. (A and B) HLA-DRB1*0301 transgenic mice (n = 15) were immunized with adjuvanted SLA/LP on days 0 and 21. (A) SLA/LP-specific antibody responses were quantified at the indicated time points and (B) studied for isotype distribution on day 42. The dashed horizontal line indicates the cut-off level of positivity. (C) Fine mapping of the antibody response of immunized HLA-DRB1*0301 transgenic (filled bars) and C57BL/6 mice (open bars). Mean and SD of the results of 3 mice are shown in B and C. A P value of less than .05 (Student t test) was considered significant.
Gastroenterology  , DOI: ( /j.gastro )
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4. Figure 3
Cellular immune response of SLA/LP-immunized HLA-DRB1*0301–transgenic mice. (A) SLA/LP dose-dependent proliferation and (B) IL-2 ELISpot using spleen cells from SLA/LP-immunized HLA-DRB1*0301–transgenic mice. (A and B) Left, results (mean of triplicate values) from representative SLA/LP-immunized mice; right, mean and SD of 3 mice per group. *Confluent spots too numerous to count. (C) Fine mapping of the immune response of SLA/LP-immunized mice (n = 11) using overlapping peptides. Filled bars indicate responses higher than the mean plus 3 SD of the medium control. Immunogenic regions for which T-cell hybrids could be established from either SLA/LP protein-immunized mice (n = 11) or peptide-immunized mice (n = 5 per peptide) are indicated by filled horizontal bars.
Gastroenterology  , DOI: ( /j.gastro )
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5. Figure 4
Fine mapping of epitope SLA/LP373–386 NRLDRCLKAVRKER and SLA/LP186–197 LIQQGARVGRID. CD4+ T-cell hybrids were stimulated with 10 μg/mL indicated truncated peptides to determine the (A and C) optimal epitopes (underlined), which were shown to be (B and D) HLA-DR–restricted.
Gastroenterology  , DOI: ( /j.gastro )
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6. Figure 5
Verification of the DRB1*0301-SLA/LP373–386 tetramer and DRB1*0301-SLA/LP184–198 tetramer. (A) DRB1*0301-SLA/LP373–386 tetramer+ and (B) DRB1*0301-SLA/LP184–198 tetramer+ CD4+ T cells were detectable in spleens of SLA/LP-immunized, but not SLA/LP-nonimmunized, DRB1*0301-transgenic mice. CD62L and CD11a expression were compared on tetramer+ and tetramer− CD4+ T cells. Numbers indicate the percentage of positive cells.
Gastroenterology  , DOI: ( /j.gastro )
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7. Figure 6
Detection of tetramer+ SLA/LP-specific CD4+ T cells in human beings. (A) Gating strategy. After exclusion of doublets (left graph), gates were set on lymphocytes based on forward and side scatter (second graph from left), and on CD3+ cells after exclusion of CD8+ T cells, CD16+ monocytes, and CD19+ B cells (second gate from right). Staining of (B) CD4+ tetramer+ T cells ex vivo and (C) after enrichment with anti-PE–coupled magnetic beads. PBMCs were carboxyfluorescein diacetate succinimidylester–labeled and stimulated with either the (D) SLA/LP373–383 peptide or an (E) DRB1*0301-restricted HCV peptide. Proliferation of DRB1*0301-SLA/LP373–386+ and DRB1*0301-SLA/LP373–386− cells were compared. Numbers indicate the percentage of events in the respective gate or quadrant.
Gastroenterology  , DOI: ( /j.gastro )
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8. Figure 7
Detection of interferon-γ+ SLA/LP-specific CD4+ T cells in human beings. (A and B) ELISpot images of tetramer-enriched CD4+ T cells of AIH patients. (B) IFN-γ producing, SLA/LP–specific cells are detected ex vivo (open symbols) or after antigen-nonspecific in vitro expansion (filled symbols) in tetramer-enriched CD4 T-cell populations of HLA-DRB1*0301–positive patients with anti-SLA/LP positive AIH but not HLA-DRB1*0301–negative patients with anti-SLA/LP–positive AIH and not HLA-DRB1*0301–positive controls without anti-SLA/LP–positive AIH. Horizontal lines indicate the mean. Please note that the number of interferon-γ–producing cells in this panel is calculated for 106 cells, but that the number of cells in the ELISpot wells in A was lower.
Gastroenterology  , DOI: ( /j.gastro )
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